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Am J Physiol Cell Physiol 289: C1522-C1534, 2005. First published July 27, 2005; doi:10.1152/ajpcell.00126.2005
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MUSCLE CELL BIOLOGY AND CELL MOTILITY

Divergent transcriptional enhancer factor-1 regulates the cardiac troponin T promoter

Anthony Azakie,1,2 Lauren LaMont,1,2 Jeffrey R. Fineman,1,2 and Youping He1

Departments of Surgery1 and Pediatrics,2 University of California, San Francisco, San Francisco, California

Submitted 21 March 2005 ; accepted in final form 22 July 2005

MCAT elements are essential for cardiac gene expression during development. Avian transcriptional enhancer factor-1 (TEF-1) proteins are muscle-enriched and contribute to MCAT binding activities. However, direct activation of MCAT-driven promoters by TEF-1-related proteins has not been uniformly achieved. Divergent TEF (DTEF)-1 is a unique member of the TEF-1 multigene family with abundant transcripts in the heart but not in skeletal muscle. Herein we show that DTEF-1 proteins are highly expressed in the heart. Protein expression is activated at very early stages of chick embryogenesis (Hamburger-Hamilton stage 4, 16–18 h), after which DTEF-1 becomes abundant in the sinus venosus and is expressed in the trabeculated ventricular myocardium and ventricular outflow tracts. By chromatin immunoprecipitation, DTEF-1 interacts with the cardiac troponin T (cTnT) promoter in vivo. DTEF-1 also interacts with MEF- 2 by coimmunoprecipitation and independently or cooperatively (with MEF-2) trans-activates the cTnT promoter. DTEF-1 isoforms do not activate the cTnT promoter in fibroblasts or skeletal muscle. DTEF-1 expression occurs very early in chick embryogenesis (16–18 h), preceding sarcomeric protein expression, and it activates cardiac promoters. As such, DTEF-1 may be an early marker of the myocardial phenotype. DTEF-1 trans-activates the cTnT promoter in a tissue-specific fashion independent of AT-rich, MEF-2, or GATA sites. The observed spatial pattern suggests decreasing levels of expression from the cardiac inlet to the ventricular outflow tracts, which may mark a cardiogenic or differentiation pathway that parallels the direction of flow through the developing chick heart.

cardiac differentiation; MCAT sites; monocyte enhancer factor-2; cell specification



Address for reprint requests and other correspondence: A. Azakie, 513 Parnassus Ave., Rm. S-549, Box 0117, Univ. of California San Francisco, San Francisco, CA 94143




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